JP2013040782A - Optical measurement apparatus and chip life determination method - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a chip life determination method for urging a user to replace a chip.SOLUTION: An optical measurement apparatus includes: a light irradiation part for applying light to a sample passing in a flow channel formed in an attachable detachable chip; a light detection part for detecting optical information generated from the sample due to light irradiation from the light irradiation part; and a determination part for determining replacement time of the chip based on the optical information detected by the light detection part. The chip life determination method includes the steps of: applying light to the sample passing in the flow channel formed in the attachable/detachable chip; detecting the optical information generated from the sample due to the light irradiation from the light irradiation part; and determining the replacement time of the chip based on the optical information detected by the light detection part.

Description

  The present disclosure relates to an optical measurement device and a chip life determination method. More specifically, the present invention relates to an optical measurement apparatus including a determination unit that determines the life of a chip by optically detecting a sample flowing through a flow path of a detachable chip. The present disclosure also relates to a chip life determination method for determining the life of a chip by optically detecting a sample flowing through the flow path of a detachable chip.

  In recent years, with the development of analytical methods, biological microparticles such as cells and microorganisms, microparticles such as microbeads are passed through the flow path, and the microparticles are individually measured or measured in the flow process. A technique for analyzing and sorting microparticles and an optical measuring device using the technique are being developed.

  For example, as a typical example of a technique for analyzing or sorting microparticles using such a flow channel, a technical improvement of an analysis technique called flow cytometry is rapidly progressing. Furthermore, removable microchips have been used in flow cytometry. For example, a microchip is known that includes a flow path through which a sheath liquid can flow and a microtube for introducing a sample liquid into a sheath liquid laminar flow that passes through the flow path (for example, Patent Documents). 1).

  There are various analysis and fractionation techniques for microparticles in the flow channel such as flow cytometry, such as medical field, drug discovery field, clinical laboratory field, food field, agricultural field, engineering field, forensic field, criminal field, etc. Widely used in various fields. Particularly in the medical field, it plays an important role in pathology, tumor immunology, transplantation, genetics, regenerative medicine, chemotherapy and the like.

  In the optical measuring apparatus as described above, a removable microchip, particularly a disposable after a predetermined period of use, for the purpose of improving the measurement accuracy and reducing the cleaning of the apparatus to improve the working efficiency. In many cases, a chip based on the assumption is used. However, in practice, the user actually uses the chip for a long time or reuses it after cleaning. In addition, when the user determines the lifetime of the detachable chip, since it is based on the experience and intuition of each user, there is a problem that the measurement accuracy of the optical measuring device is lowered or trouble is likely to occur. .

JP 2010-54492 A

Thus, there is a demand for a method and apparatus that can prompt chip replacement without leaving the chip replacement time (hereinafter also referred to as “chip life”) to the experience and intuition of individual users.
Therefore, in view of such a situation, the present disclosure mainly aims to provide an optical measurement device and a chip life determination method that prompt the user to replace the chip.

  The present disclosure relates to a light irradiation unit that irradiates light to a sample flowing through a flow path in a detachable chip, and light that detects optical information emitted from the sample by light irradiation by the light irradiation unit An optical measurement device is provided that includes a detection unit and a determination unit that determines the replacement time of a chip based on optical information detected by the light detection unit. As a result, it is not necessary to leave the judgment of the chip life (replacement time) to the experience and intuition of each user, and it is possible to stably perform the life judgment for each chip with little variation. In addition, the condition setting can be freely changed as follows according to the user's request such as the purpose of measurement and the object.

Further, it is preferable to provide a chip information recognition unit that recognizes chip information from the identifier, and to determine the replacement time of the chip based on the optical information and / or the chip information. Thereby, it is possible to grasp the usage state of the chip more accurately.
It is preferable that the optical information is obtained by sorting with a threshold value. This makes it possible to remove noise and the like that are likely to cause a determination error when determining the chip life (replacement time). For this reason, it becomes easy to perform the life determination for each chip without variation.
When the determination unit determines that the number of samples calculated based on the optical information has reached a certain value of the maximum count number, it is preferable to determine that it is time to replace the chip.
When it is determined that the determination unit has measured a large amount of samples at a time based on the optical information, it is preferable that the determination unit determine that it is time to replace the chip after a predetermined time has elapsed.
When the determination unit determines that the sample size calculated based on the optical information has reached a certain value of the maximum integration number of the sample size, it is preferable to determine that it is time to replace the chip. is there.

  Further, the present disclosure provides a procedure for irradiating light to a sample flowing through a flow path in a detachable chip, and a procedure for detecting optical information emitted from the sample by light irradiation by the light irradiation unit. And a procedure for determining the replacement time of the chip based on the optical information detected by the light detection unit.

  Here, the “sample” in the present disclosure refers to biologically relevant microparticles such as cells, microorganisms, liposomes, DNA, and proteins, or synthetic particles such as latex particles, gel particles, and industrial particles. Include all possible substances.

  According to the present disclosure, it is possible to promote the replacement of a chip without leaving the chip life to the experience and intuition of each user.

1 is a schematic conceptual diagram schematically showing a first embodiment of an optical measuring device 1 according to the present disclosure. 1 is a schematic conceptual diagram schematically showing a first embodiment of an optical measuring device 1 according to the present disclosure. It is a figure of flow 1 of a chip life judging method concerning this indication. It is a figure of flow 2 of a chip life judging method concerning this indication. It is a figure of flow 3 of the chip life judging method concerning this indication. It is a figure of flow 4 of a chip life judging method concerning this indication. It is a figure of flow 5 of a chip life judging method concerning this indication. It is a drawing substitute graph which shows the example (in the case of two particles, three) of the state of optical information (pulse signal etc.) when two samples S flow in large quantities at once. It is a figure of flow 1 of a chip life judging method concerning this indication. It is a block diagram which shows the structure of a determination part. It is a figure of the graph which shows the number of accumulation events (pulse signal etc.) when 10,000 event per secx60secx5 life is made into 1 chip.

  DESCRIPTION OF EXEMPLARY EMBODIMENTS Hereinafter, preferred embodiments for carrying out the invention will be described with reference to the drawings. In addition, embodiment described below shows an example of typical embodiment of this invention, and, thereby, the range of this invention is not interpreted narrowly. The description will be given in the following order.

1. Optical measuring device (1) Flow path (2) Light irradiation part (3) Light detection part (4) Electrical signal conversion part / AD conversion part (5) Chip information recognition part (6) Determination part 2. Chip life determination method Flow cytometer

<1. Optical measuring device>
1 and 2 are schematic conceptual views schematically showing an embodiment of an optical measuring device 1 according to the present disclosure.
The optical measurement device 1 according to the present disclosure is roughly provided with at least a light irradiation unit 11, a light detection unit 12, and a determination unit 13.
The optical measuring device 1 can be mounted with a chip (substrate T) having a removable flow path 2. Further, when the chip (substrate T) is accompanied by the identifier I, a chip information recognition unit 14 that recognizes the chip information from the identifier I may be provided.
Moreover, you may provide the electrical signal conversion part (not shown) which converts optical information into an electrical signal (light pulse), and the AD conversion part (not shown) which performs analog-digital conversion. Note that the optical measurement apparatus of the present disclosure may further include a unit that adjusts the laminar flow of the flow path 2 in the chip, a temperature control unit, a sorting unit, a control unit that controls the function of each unit, and the like. Good. Further, the control unit may perform processing performed by a determination unit, a chip information recognition unit, an electrical signal conversion unit, an AD conversion unit, and the like.

According to the present disclosure, it is possible to prompt the user to replace the chip without leaving the determination of the chip replacement time (chip life) to the experience and intuition of each user. That is, the present disclosure automatically calculates and predicts the chip usage status such as the usage frequency and usage period of the chip, and prompts the user to replace the chip that has been used or will be used for a certain amount or period. Is possible.
And since there is little variation in the lifetime judgment for each chip, it is possible to stably and efficiently replace the chip. Further, it is possible to freely change the condition setting of the chip life according to the user's request such as the purpose and object of measurement. It is also possible to determine the chip life in real time. Thereby, it is also possible to grasp the usage state of the chip more accurately. In addition, the running cost of optical measurement can be reduced.

(1) Chip The optical measuring device 1 of the present disclosure can be mounted with a chip (substrate T) having a removable flow path 2. For example, the mounting position of the chip may be arranged so that the light irradiation unit 11 and the light detection unit 12 face each other. For example, the vertical type and horizontal type chips can be mounted.

Further, the chip is preferably accompanied by an identifier I from which chip information can be acquired (see FIG. 2 as an example). The position of the identifier I is not particularly limited. For example, the identifier I may be given to the package of the chip; the identifier I may be attached to the chip; the identifier I may be present in the chip body or may be removable.
This makes it easy to grasp the chip usage state such as the usage frequency and usage period for each chip, so that the replacement time for each chip can be determined more accurately.
Here, examples of the identifier I include a barcode, a data matrix barcode, a radio frequency (RFID), a marker, a character, a shape (unevenness, protrusion, notch, groove, etc.) and the like. From this identifier I, chip information can be acquired via the chip information recognition unit.
The chip information includes information related to the lifetime (replacement time) of the chip. Examples of the chip information include distinction for each chip, chip usage frequency, chip usage period, operation status of the device when the chip is used, usage status of the chip at that time, and the like.
The storage unit for storing the chip information is not particularly limited, and examples thereof include a storage unit existing in a chip identifier I, a device to be used, a network, and the like.

Then, the sample S flows through the flow channel 2, and the sample S is irradiated with light by a light irradiation unit 11 described later at a predetermined portion of the flow channel 2, and the sample S is detected by the light detection unit 12. Various optical information can be obtained.
Further, the channel width, the channel depth, and the channel cross-sectional shape of the channel 2 are not particularly limited as long as the channel can be formed, and can be freely designed. Examples of the channel 2 include a micro channel having a channel width of 1 mm or less (more specifically, a channel width of about 10 μm to 1 mm).

In addition, when employ | adopting the flow path 2 formed in the chip | tip (substrate T), it is preferable to form the surface of the flow path 2 with a transparent material.
As the chip, for example, as shown in FIG. 1, a configuration in which the flow path 2 is formed by a plurality of substrates is preferable.
Further, as shown in FIG. 2, a chip formed so as to join the flow path 2 by sandwiching the flow path for sample liquid injection at the substantially center between the flow paths for two sheath liquid injections may be used. Thereby, a sheath liquid laminar flow and a sample flow laminar flow are formed, and the sample S flows through the flow path 2. In the case of such a chip, after the chip is set in the optical measuring device, only the sheath liquid is first flowed, and then the sample liquid is also flowed to start the optical measurement.

  The substrate used as the chip can be formed by wet etching, injection molding, cutting, or the like of the substrate layer. The material of the substrate is not particularly limited, and can be appropriately selected in consideration of a detection method, processability, durability, and the like. The material may be appropriately selected depending on the desired optical analysis as a material having heat resistance or light transmittance, such as glass and various plastics (polypropylene, polycarbonate, cycloolefin polymer, polydimethylsiloxane, etc.), etc. Is mentioned.

(2) Light irradiation part The light irradiation part 11 of this indication irradiates light with respect to the sample S currently flowing through the flow path 2 of the chip | tip which can be attached or detached.
The light irradiation unit 11 may use a light source corresponding to a desired type of light. The type of light emitted from the light source is not particularly limited, but in order to reliably generate fluorescence and scattered light from the sample S, it is desirable that the light has a constant light direction, wavelength, and light intensity. As an example, a laser, LED, etc. can be mentioned. In the case of using a laser, the type is not particularly limited, but one or more of argon ion (Ar) laser, helium-neon (He-Ne) laser, second (dye) laser, krypton (Cr) laser, etc. Can be combined freely.

(3) Light Detection Unit The light detection unit of the present disclosure detects optical information emitted from the sample S by light irradiation by the light irradiation unit 11.
The light detection unit 12 is not particularly limited as long as it can detect optical information, and a known light detector can be freely selected and employed.
For example, fluorescence measuring instrument, scattered light measuring instrument, transmitted light measuring instrument, reflected light measuring instrument, diffracted light measuring instrument, ultraviolet spectroscopic measuring instrument, infrared spectroscopic measuring instrument, Raman spectroscopic measuring instrument, FRET measuring instrument, FISH measuring instrument, etc. Is mentioned. In addition, other various spectrum measuring devices, a so-called multichannel photodetector in which a plurality of photodetectors are arranged in an array, and the like can be given. These can be used alone or in combination of two or more. In this way, it is possible to obtain optical information generated from the sample S by combining the light irradiation unit 11 and the light detection unit 12.

  Further, the installation location of the light detection unit 12 is not particularly limited as long as optical information emitted from the sample S can be detected, and can be freely designed. For example, as shown in FIGS. 1 and 2, it may be arranged on the opposite side of the light irradiation unit 11 with the flow channel 2 interposed therebetween.

(4) Electrical Signal Conversion Unit / AD Conversion Unit The chip life determination device 1 of the present disclosure may further include an electrical signal conversion unit (not shown) that converts the optical information into an electrical signal. . This makes it possible to obtain a peak (light pulse) due to signal intensity.
The converted electrical signal can be further AD converted by an analog-digital converter (not shown). Thereafter, based on this digital data, a numerical calculation processor or the like can extract a histogram with an analysis computer and software and perform analysis (digital waveform processing or the like).
The optical information detected by the light detection unit 12 can be converted into a pulse (pulse shape) as shown in FIGS. 1 and 2, for example, by a digital waveform processing unit (not shown). Based on the pulse shape, the baseline, and the threshold value, the peak height, the peak width, and the peak area / volume calculated from the height and width can be calculated. Note that the threshold value can be appropriately set and changed by a determination unit described later.
In this way, data such as the number of light pulses, the number of light pulses having a height exceeding a threshold, and the height, width, area, etc. of each light pulse can be obtained based on optical information by a CPU, a processor, or the like. It is.

(5) Chip Information Recognition Unit It is preferable that the optical measurement device 1 of the present disclosure further includes a chip information recognition unit 14. The chip information identification unit 14 may be connected to the optical measurement device 1 via a network, a communication cable, or the like. The chip information recognition unit 14 is a device that can recognize the identifier I as chip information by machine recognition or the like (see, for example, FIG. 2). The chip information recognition part 14 should just be arrange | positioned so that the identifier I can be recognized.
As the recognition method, for example, an identifier I (for example, a barcode, a character, a three-dimensional shape, etc.) is imaged and read by a chip information recognition unit 14 (for example, a barcode reader, a CCD camera, etc.), and the identifier Image data is obtained.
As an example, the identifier data assigned to the chip package may be acquired by an image recognition device such as a CCD camera connected to or incorporated in a personal computer or the like. Further, when the chip is mounted on the optical measuring device, the identifier I provided on the chip may be acquired by an image recognition device or the like.
Further, when the shape of a notch or the like formed on the chip is contacted or joined to the sensor or the like of the chip information recognition unit 14, the identifier is recognized and the data is obtained.
Each data of the obtained identifier is converted by a CPU or the like according to a certain rule, and becomes chip information such as numbers, characters and symbols.
Further, as the recognition method, for example, when the identifier I provided on the substrate is a radio frequency (RFID) or the like, the chip information stored in the identifier is obtained by the chip information recognition unit 14 having a wireless function or the like. It is possible.

The chip information recognition unit 14 collates the chip information obtained from the chip identifier with each data of chip information stored in the storage unit 28 or the like to be described later. It is possible to determine whether or not.
Further, chip information, for example, information on the usage state (usage frequency, usage period, etc.) of each chip can be stored in a storage unit provided in an optical measurement device, a network, or the like.
Thereby, even when the power of the apparatus is turned off, it is possible to determine the replacement time of the chip after restarting. In addition, when a network system or the like is constructed using a plurality of devices, it is possible to determine the replacement time of a chip even for a chip used for another device. It is also possible to update and store new chip information obtained during measurement.

(6) Determination Unit The determination unit 13 of the present disclosure determines the replacement time of the chip based on the optical information detected by the light detection unit 12 (see, for example, FIGS. 3 to 8).

  FIG. 9 shows an example of a schematic configuration of the determination unit 13. The determination unit 13 has at least a CPU (Central Processing Unit) 21. The determination unit 13 can be configured by connecting various types of hardware to the CPU via the bus 22. In addition, these CPU and various hardware may utilize various hardware resources with which the optical measuring device is equipped.

  As the hardware, at least a ROM (Read Only Memory) 23, a RAM (Random Access Memory) 24 serving as a work memory of the CPU 21, and an interface 25 are employed. In this embodiment, an input unit 26, a display unit 27, and a storage unit 28 that can input commands according to user operations are also employed.

The storable hardware such as the ROM 23 and the storage unit 28 stores a program (hereinafter also referred to as “chip life determination program”) for executing processing of a <chip life determination method> described later. The light irradiation unit 11, the light detection unit 12, the signal conversion unit, the AD conversion unit, and the chip information identification unit 14 are connected to the interface 25.
When the CPU 21 is given an instruction for determining the chip life (replacement time) by the operation input unit 26 or chip insertion, the CPU 21 develops a chip life determination program stored in the ROM 23 or the like in the RAM 24, and the wavelength analysis unit or individual It can function as an identification processing unit or the like. The wavelength analysis unit can process optical information and analyze the number of peak vertices, width, area, volume, and the like of an optical pulse. The chip information recognition unit 14 can perform chip information machine recognition processing, conversion processing from identifier data to chip information, and the like.
The CPU 21 can also control the apparatus based on optical information and chip information.
Further, the CPU 21 can store the information data in the storage unit 28 and use it for calculation or the like as appropriate.

Then, the determination unit 13 determines the chip life (chip replacement time) according to <Chip life determination method> described later. Furthermore, the optical information is preferably converted into digital data.
In general, the chip may be used for a long time or reused after being washed. In such a case, particles or the like are likely to adhere to the flow path of the chip, and the flow path is likely to be clogged. In general, the sample flow tends to be stagnation. In particular, it is likely to occur at a change point of the flow path (for example, a long and narrow flow path).
On the other hand, the chip life determination of the present disclosure allows the chip life before troubles such as clogging or stagnation due to the sample or buffer solution in the flow path without leaving the life of the chip to the experience and intuition of individual users. It is possible to prompt the user to replace the chip.
In addition, it is desirable that threshold data set with various conditions is stored in advance in the storage unit 28 or the like. Then, analog or digital data of the obtained optical information can be selected based on a threshold value. As a result, it is possible to determine the replacement time of the chip more accurately and with less variation.

<2. Chip life judgment method>
The chip life determination method (determination procedure) of the present disclosure includes at least a determination procedure for determining the replacement time of the chip based on optical information derived from the sample S flowing through the flow path 2 in the removable chip. Is included.
More preferably, it includes a light irradiation procedure for irradiating the sample S passing through the flow path 2 in the detachable chip with light. In addition, a light detection procedure in which optical information emitted from the sample S by light irradiation by the light irradiation unit 11 is detected by the light detection unit 12 is included.

The following determination method is mentioned as a more preferable determination method of the chip life.
For example, there is a method for determining a chip life (replacement time) including a procedure in which one or more selected from the following (a), (b) and (c) are combined.
Specifically, (a) when it is determined that the number of samples S calculated based on the optical information has reached a certain value of the maximum count number, it is determined that the life of the chip (replacement time) (see FIG. 3).
Further, (b) when it is determined that a large number (a plurality) of samples S are measured at a time based on the optical information, it is determined that the lifetime (replacement time) of the chip after a predetermined time has elapsed (see FIG. 4). Can be mentioned.
(C) When it is determined that the size of the sample S calculated based on the optical information has reached a certain value of the maximum integrated number of the size of the sample S, it is determined that the life of the chip (replacement time) (See FIG. 5).

When the flow 1 shown in FIG. 3 is used, (a) when it is determined that the number of samples S calculated based on the optical information has reached a certain value of the maximum count number, it is determined that it is time to replace the chip. This will be explained.
The determination unit 13 uses, as a reference (hereinafter also referred to as “one peak reference”), one peak derived from optical information emitted from one sample S (cell) in the flow path 2 of the chip after the measurement is started. One peak vertex is determined to be 1 count. As the one-peak reference, an average value of a plurality of peaks to be measured can be used. The count number may be an integer.
Furthermore, for example, the following conditions (A) and (B) may be set as appropriate. (A) As shown in FIG. 8, the peak of the measured peak occurs and a peak having a plurality of peak vertices is selected. In this case, the number of peak vertices of the peak crack is counted. Also, (a) a peak whose peak is higher than the height of “one peak standard” is selected. Divide the height of the selected high peak by the height of the “one peak reference”. The quotient at this time is counted as the number of peak vertices.

Furthermore, it is desirable to set a threshold value for selecting peaks by height in the determination unit 13. If the peak is higher than this threshold, it is counted, and if it is lower than the threshold, it is not counted. Thereby, it is possible to reduce the error of the peak count due to the influence of noise or the like.
The situation where the measured peak exceeds a set threshold is also referred to as an “event”.

The determination unit 13 preferably stores measurement data such as the above-described count number (value), measurement time, and signal intensity in the storage unit 28, the RAM 24, and the like. Based on this data, it is also possible to calculate the number of events (event per sec: eps) per time (second: Sec).
Then, when the total count value of the number of peak vertices (cell count number) reaches a certain value (upper limit) of the maximum count number stored in advance in the storage unit 28 or the like (YES), the determination unit 13 Determines that it is time to replace the chip and causes the display unit 27 to display “Warning”. This warning display prompts the user to replace the chip.
Here, the “maximum count number” is a value at which the possibility of clogging, a decrease in detection accuracy, or the like in the flow path becomes extremely high when the count value is exceeded.
The “display” may be anything that can be understood by visual, auditory, tactile, or the like, and may be light, sound, vibration, or the like in addition to an image. Further, it may be displayed on a network terminal or the like via a network. The means and method are not particularly limited.
The determination unit 13 can also predict the arrival time to the maximum number of counts based on the eps and display the arrival prediction time and the like together with the “pre-warning” before the warning.

On the other hand, if it is determined that the total count value has not yet reached the constant value of the maximum count value (for example, the number of events> 1,000,000) (NO), it is further determined whether or not “measuring”.
If it is determined that the measurement is in progress (YES), the optical pulse count is continued. When it is determined that it is not “measuring” (NO), “new measurement” is started with a new sample.
At this time, the number of peak vertices so far and the obtained data such as the eps are stored in the storage unit 28 or the like. As a result, the cumulative number of peak vertices of the chip (for example, the cumulative number of events) can be used even after “new measurement”. Therefore, it is possible to display a warning with higher accuracy.

Using the flow 2 shown in FIG. 4, when it is determined that (b) a plurality of samples S (cells, etc.) have been measured in large quantities at once based on the optical information, it is determined that the chip replacement time has elapsed after a certain period of time. I will explain what to do. The same parts as those in the flow 1 described above are omitted as appropriate.
The determination unit 13 determines whether or not a plurality of samples S have passed through the measurement part in a large amount at once in the flow path of the chip 2 after the start of measurement. When a large amount of sample S passes at once, there is a high possibility that the concentration of the sample solution being measured is high, and the possibility of clogging the flow path is increasing.

When the determination unit 13 determines that the measured peak size is “large peak” as compared to “one sample (cell) one peak” (the one peak reference), the plurality of samples S are It is determined that a predetermined amount of the passage has been passed in large quantities at a time.
Here, the comparison between the measured peak and the one-peak standard may be performed based on the peak shape (height, width, area / volume). Among these, peak breakage and broad peak due to data processing speed and mass passage are included. Considering the peak area, it is preferable.
At this time, it is desirable to set a threshold value for selecting the measured peak by shape (height, width, area / volume, etc.) in the determination unit 13.
For example, by increasing the threshold setting, it is possible to display as an “emergency warning” that a very large number of samples S have flowed at once. On the other hand, by lowering the threshold setting, it is possible to display “warning” even if a small number of samples S flow at once. Thereby, the warning level can be changed and displayed according to the number of samples S passing at a time, so that troubles such as clogging can be easily avoided.
A plurality of threshold values may be set. It is possible to set a plurality of warning levels corresponding to a plurality of set threshold values and display warning levels according to the levels.
The determination unit 13 may set the number of determinations when it is determined that a large amount of the sample S has been measured at one time. When the set number of determinations is exceeded, an “emergency warning” can be displayed.

In this way, if it is determined that a large amount of sample S has been measured at one time ("large peak") (YES), the possibility of clogging of the flow path increases after a certain period of time, so it is determined that it is time to replace the chip. Then, “Warning” is displayed on the display unit.
If it is determined that a large amount of sample S has not been measured at once (NO), it is determined whether or not it is “measuring”. If it is determined that the measurement is in progress (YES), the optical pulse count is continued. When it is determined that it is not “measuring” (NO), “new measurement” is started with a new sample.

The starting point of the “certain time” may be the starting point when it is determined as a “large peak”. In addition, the width of “fixed time” can be set in consideration of the size of “big peak”. For example, when “big peak” is larger, the width of “fixed time” is set short. If the “large peak” is smaller, the “certain time” may be set longer. These starting point and width settings may be stored in the storage unit 28 in advance.
It should be noted that a “warning” may be displayed immediately without causing “a certain period of time”. Alternatively, “pre-warning” may be displayed when the “large peak” is determined, and then “warning” may be displayed after a predetermined time has elapsed.

When the flow 3 shown in FIG. 5 is used, (c) when it is determined that the size of the sample S calculated based on the optical information has reached a certain value of the maximum integration number of the size of the sample S, the chip The determination of the replacement time will be described. The same parts as those in the above flow 1 are omitted.
The determination unit 13 can determine the size of the cell flowing in the flow path 2 of the chip after the start of measurement from the peak area as described above. The determination unit 13 calculates the peak area, and stores the accumulated area number in the storage unit 28 or the like. Note that “peak area” may be determined instead of “peak volume”.
At this time, it is desirable to set a threshold value for selecting a peak by area (volume) in the determination unit 13. By providing the threshold value, it is possible to remove noise and obtain a more accurate peak area.
Here, the “maximum integration number” is a maximum value when optical pulses or the like are integrated, and is a value that extremely increases the possibility of clogging or a decrease in detection accuracy in the flow path.

  When the total peak area reaches a certain value of the maximum integration number (upper limit value) of peak area integration stored in advance in the storage unit 28 or the like (YES), the determination unit 13 replaces the chip. The time is determined and “warning” is displayed on the display unit 27. This warning display prompts the user to replace the chip. If it is determined that the maximum integrated number of peak areas has not been reached (NO), it is determined whether or not “measuring”. If it is determined that the measurement is in progress (YES), the optical pulse count is continued. If it is determined that the measurement is not in progress (NO), “new measurement” is started on the specimen including the new sample.

Further, for example, it is possible to combine two or more of the above-described (a), (b), and (c). The order of (a), (b) and (c) may be changed as appropriate. When two or more of (a) to (c) are applicable, it is possible to determine that it is time to replace the chip.
Further, when any one of (a) to (c) is applicable, it is possible to determine that it is time to replace the chip (see, for example, FIG. 6), and to prevent clogging of the flow path. It becomes possible.

The determination unit 13 can calculate the number of peak vertices, peak height, peak width, peak area (volume), and the like based on the optical information. Furthermore, these may be integrated and these data may be stored in the storage unit 28 or the like. The total count value of the number of vertices of the predetermined peak, the predetermined peak area, and the upper limit value of the predetermined peak area integration are stored in the storage unit 28.
Then, the determination unit 13 determines whether the total count value of the number of peak vertices is equal to or greater than the total value of predetermined peak vertices (YES) or not (NO). If the value is equal to or greater than the predetermined value, “warning” is displayed.

If it is not equal to or greater than the total value of the predetermined peak vertices (NO), the determination unit 13 further determines whether or not a large amount of the sample S has been measured at one time. Further, it is determined whether or not (YES) or not (NO) over a predetermined peak area (volume). “Warning” is displayed when the peak area (volume) is not less than a predetermined value.
The determination unit 13 may display “preliminary warning” when the peak area (volume) is not equal to or greater than (NO). Furthermore, the determination part 13 determines whether it is more than the upper limit (YES) of predetermined peak area integration (NO).
If it is equal to or greater than a predetermined peak area integration upper limit (YES), “warning” is displayed. If it is not equal to or greater than the upper limit (NO), it is further determined whether or not “measuring”. If it is determined that the measurement is in progress (YES), the optical pulse count is continued. If it is determined that the measurement is not in progress (NO), “new measurement” is started on the specimen including the new sample.

Further, by using the identifier I and the chip information recognition unit 14, it is possible to measure the time elapsed since the chip was mounted on the apparatus. If the chip mounting time is equal to or longer than the predetermined mounting time, it is determined that it is time to replace the chip, and “warning” can be displayed on the display unit.
As an example, when a chip is inserted, the chip information from the identifier I is referred to as the chip information stored in the storage unit 28. As a result of the reference, when the device is mounted for the first time (YES), counting of optical pulses is started. On the other hand, if the device is mounted on the device one or more times (NO), the optical pulse count is not started, it is determined that the chip is to be replaced, and “warning” is displayed on the display unit. Thereby, it is possible to prevent the chip from being used twice.

As an example, as shown in FIG. 7, the determination unit 13 checks the chip information (chip ID and the like) after the chip is inserted, and counts the optical pulse if the predetermined time has not elapsed (NO). Start. If a predetermined time has elapsed after the chip insertion (YES), the optical pulse count is not started, it is determined that the chip is to be replaced, and “warning” is displayed on the display unit. The mounting time after chip insertion may be stored in the storage unit 28 over time.
Whether the mounting time after chip insertion is less than the predetermined time has elapsed may be determined by the determination unit 13 or may be determined by the chip information recognition unit 14. The result of the chip information recognition unit 14 may be transmitted to the determination unit 13, and the determination unit 13 may display “warning” or determine “under measurement”.

<3. Flow cytometer>
Furthermore, the optical measurement apparatus according to the present disclosure can be suitably used for a flow cytometer by utilizing its high accuracy.
“Flow cytometry” refers to the flow of fluorescent particles and scattered light emitted from each microparticle by pouring the microparticles to be analyzed in a state of being aligned in a fluid and irradiating the microparticles with laser light or the like. This is an analysis method for analyzing and sorting microparticles (sample S) by detecting. Flow cytometry processes can be broadly classified into the following (1) water flow system, (2) optical system, (3) electrical / analysis system, and (4) preparative system.

(1) Water flow system In the water flow system, fine particles to be analyzed are aligned in a flow cell (flow channel). More specifically, the sheath flow is introduced into the flow cell at a constant flow rate, and in this state, a sample flow containing microparticles is slowly injected into the center of the flow cell. At this time, due to the principle of laminar flow, the respective flows are not mixed with each other, and a laminar flow (laminar flow) is formed. Then, the inflow amounts of the sheath flow and the sample flow are adjusted according to the size of the microparticles to be analyzed, and the microparticles are allowed to flow in an aligned state.

(2) Optical system In an optical system, light, such as a laser, is irradiated to fine particles to be analyzed, and fluorescence and scattered light emitted from the fine particles are detected. In the water flow system (1), the fine particles are flowed through the laser irradiation section in a state where the fine particles are aligned one by one, and each time the fine particles pass, fluorescence and scattered light emitted from the fine particles are emitted. Each parameter is detected using an optical detector and the characteristics of each microparticle are analyzed.

(3) Electricity / analysis system The electricity / analysis system converts optical information detected by the optical system into an electric signal (voltage pulse). The converted electrical signal is subjected to analog-digital conversion, and a histogram is extracted and analyzed using an analysis computer and software based on this data.
For example, in a pulse detection system, fluorescence or scattered light generated when a microparticle crosses a laser is detected as an electric pulse, and analysis is performed by analyzing a pulse height, a pulse width, a pulse area, and the like.

(4) Preparative system The preparative system separates and collects fine particles that have been measured. As a typical sorting method, a positive or negative charge is added to the microparticles for which measurement has been completed, the flow cell is sandwiched between two deflecting plates having a potential difference, and the charged microparticles are selected according to the charge. There is a method of sorting by being drawn to the deflection plate.

In addition, this technique can also take the following structures.
[1] A light irradiation unit that irradiates light to a sample flowing through a flow path in a detachable chip, and light that detects optical information emitted from the sample by light irradiation by the light irradiation unit A chip life determination apparatus comprising: a detection unit; and a determination unit that determines a chip replacement time based on optical information detected by the light detection unit.
[2] The chip life determination apparatus according to [1], further including a chip information recognition unit for identifying chip information from an identifier, and determining a chip replacement time based on the optical information and / or the chip information.
[3] The chip life determination apparatus according to [1] or [2], wherein the optical information is obtained by sorting with a threshold value.
[4] Any one of [1] to [3], wherein when the number of samples calculated based on the optical information reaches a constant value of the maximum count number, it is determined that the chip replacement time is reached. Chip life judgment device.
[5] Chip life determination according to any one of [1] to [4], in which when a large number of samples are measured at a time based on the optical information, it is determined that the chip replacement time is reached after a predetermined time has elapsed. apparatus.
[6] The above-mentioned [1] to [5], in which when the sample size calculated based on the optical information reaches a constant value of the maximum integrated number of sample sizes, it is determined that the tip is to be replaced. The chip life judging apparatus of any one of these.

[7] An optical measuring device comprising the chip life judging device according to any one of [1] to [6].

[8] A procedure for irradiating light to the sample flowing through the flow path of the detachable chip, a procedure for detecting optical information emitted from the sample by light irradiation by the light irradiation unit, and the light And a procedure for determining the replacement time of the chip based on the optical information detected by the detection unit.
[9] The chip according to [8], wherein the chip information recognition unit recognizes chip information from an identifier associated with the chip, and determines a replacement time of the chip based on the optical information and / or the chip information. Life judgment method.
[10] The chip life judging method according to the above [8] or [9], wherein the optical information is obtained by sorting with a threshold value.
[11] The method according to any one of [8] to [10], wherein when the number of samples calculated based on the optical information reaches a constant value of the maximum count number, it is determined that the chip replacement time is reached. Chip life judgment method.
[12] The chip life determination according to any one of [8] to [11], wherein when a large amount of samples are measured at a time based on the optical information, the chip replacement time is determined after a predetermined time has elapsed. Method.
[13] The above-mentioned [8] to [12], in which when the sample size calculated based on the optical information reaches a constant value of the maximum integrated number of sample sizes, it is determined that the tip is to be replaced. The chip life judging method according to any one of the above.

[14] A program for executing the chip life judging method according to any one of [8] to [13].

  FIG. 10 shows the results of the cumulative number of events (pulses) when an unused microchip is mounted on the flow cytometer and the lifetime (replacement time) is 10,000 eps × 60 sec × 5 times per chip.

  According to the present disclosure, with respect to a technique for optically detecting a sample flowing through a flow path, the lifetime of a chip including the flow path can be determined instead of the user's experience and intuition. For this reason, it is possible to maintain the analysis accuracy above a certain level. By using this technology, medical field (pathology, tumor immunology, transplantation, genetics, regenerative medicine, chemotherapy, etc.), drug discovery field, clinical laboratory field, food field, agricultural field, engineering field, forensic field , Can contribute to the improvement of analysis and analysis technology in various fields such as criminal forensics.

DESCRIPTION OF SYMBOLS 1 Optical measuring device: 11 Light irradiation part: 12 Light detection part: 13 Judgment part: 14 Fixed identification part

Claims (7)

A light irradiation unit for irradiating light to a sample flowing through a flow path in a removable chip;
A light detection unit for detecting optical information emitted from the sample by light irradiation by the light irradiation unit;
An optical measurement apparatus comprising: a determination unit that determines a replacement time of a chip based on optical information detected by the light detection unit. Furthermore, a chip information recognition unit that recognizes chip information from the identifier,
The optical measuring device according to claim 1, wherein a chip replacement time is determined based on the optical information and / or the chip information.   The optical measurement apparatus according to claim 1, wherein the optical information is obtained by sorting with a threshold value.   The optical measurement device according to claim 3, wherein the determination unit determines that it is time to replace a chip when determining that the number of samples calculated based on the optical information has reached a certain value of the maximum count.   The optical measuring device according to claim 3, wherein when the determination unit determines that a large amount of samples are measured at once based on the optical information, the determination unit determines that it is time to replace the chip after a predetermined time has elapsed.   The determination unit determines that it is time to replace a chip when determining that the sample size calculated based on the optical information has reached a certain value of the maximum integrated number of sample sizes. The optical measuring device described. A procedure for irradiating light to a sample flowing through a flow path in a detachable chip,
A procedure for detecting optical information emitted from the sample by light irradiation by the light irradiation unit;
And a procedure for determining the replacement time of the chip based on the optical information detected by the light detection unit.

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